LOCUS       sequence-466708-        5873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  chr2:43319395-43320271 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5873)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5873)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL3_THADA
FEATURES             Location/Qualifiers
     source          1..5873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1014..1135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1105..1124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1176..2828)
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     2744..2762
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      2825..2834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2864..3060)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2878..3013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    3972..4063
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(4004..4023)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    4077..4125
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4256..4711)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4393..4414)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4605..4624
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4738..4842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4843..4911,4912..5703)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5061..5080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgctgccg
      781 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
      841 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
      901 ggatccagat ccttatcgat tttaccacat ttgtagaggt tttacttgct ttaaaaaacc
      961 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1021 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1081 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1141 tctgctcgaa gcggccggcc gccccgactc tagaattaca cggcgatctt tccgcccttc
     1201 ttggccttta tgaggatctc tctgattttt cttgcgtcga gttttccggt aagacctttc
     1261 ggtacttcgt ccacaaacac aactcctccg cgcaactttt tcgcggttgt tacttgactg
     1321 gcgacgtaat ccacgatctc tttttccgtc atcgtctttc cgtgctccaa aacaacaacg
     1381 gcggcgggaa gttcaccggc gtcatcgtcg ggaagacctg cgacacctgc gtcgaagatg
     1441 ttggggtgtt ggagcaagat ggattccaat tcagcgggag ccacctgata gcctttgtac
     1501 ttaatcagag acttcaggcg gtcaacgatg aagaagtgtt cgtcttcgtc ccagtaagct
     1561 atgtctccag aatgtagcca tccatccttg tcaatcaagg cgttggtcgc ttccggattg
     1621 tttacataac cggacataat cataggacct ctcacacaca gttcgcctct ttgattaacg
     1681 cccagcgttt tcccggtatc cagatccaca accttcgctt caaaaaatgg aacaacttta
     1741 ccgaccgcgc ccggtttatc atccccctcg ggtgtaatca gaatagctga tgtagtctca
     1801 gtgagcccat atccttgcct gatacctggc agatggaacc tcttggcaac cgcttccccg
     1861 acttccttag agaggggagc gccaccagaa gcaatttcgt gtaaattaga taaatcgtat
     1921 ttgtcaatca gagtgctttt ggcgaagaag gagaataggg ttggcaccag cagcgcactt
     1981 tgaatcttgt aatcctgaag gctcctcaga aacagctctt cttcaaatct atacattaag
     2041 acgactcgaa atccacatat caaatatccg agtgtagtaa acattccaaa accgtgatgg
     2101 aatggaacaa cacttaaaat cgcagtatcc ggaatgattt gattgccaaa aataggatct
     2161 ctggcatgcg agaatctcac gcaggcagtt ctatgaggca gagcgacacc tttaggcaga
     2221 ccagtagatc cagaggagtt catgatcagt gcaattgtct tgtccctatc gaaggactct
     2281 ggcacaaaat cgtattcatt aaaaccggga ggtagatgag atgtgacgaa cgtgtacatc
     2341 gactgaaatc cctggtaatc cgttttagaa tccatgataa taattttttg gatgattggg
     2401 agcttttttt gcacgttcaa aattttttgc aacccctttt tggaaacgaa caccacggta
     2461 ggctgcgaaa tgcccatact gttgagcaat tcacgttcat tataaatgtc gttcgcgggc
     2521 gcaactgcaa ctccgataaa taacgcgccc aacaccggca taaagaattg aagagagttt
     2581 tcactgcata cgacgattct gtgatttgta ttcagcccat atcgtttcat agcttctgcc
     2641 aaccgaacgg acatttcgaa gtactcagcg taagtgatgt ccacctcgat atgtgcatct
     2701 gtaaaagcaa ttgttccagg aaccagggcg tatctcttca tagccttatg cagttgctct
     2761 ccagcggttc catcttccag cggatagaat ggcgccgggc ctttctttat gtttttggcg
     2821 tcttccatgg tggctttacc aacagtaccg gaatgccaag ctttttgcaa aagcctaggc
     2881 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     2941 ctgcataaat aaaaaaaatt agtcagcgat ggggcggaga atgggcggaa ctgggcggag
     3001 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     3061 gatcgcagat cttaagatct attcagggaa gagtgaactc tactcagata ccctctctgc
     3121 atctacgacc ccggcttttt tctattttat gtattttctt tgaaaggtca ttattttgaa
     3181 tagctgtgag ttcattctgt tccattctgt ggagataact gagctgggct catttgatgc
     3241 tgggtatttg agactctctt ctcaaatcga cccctacacc aatgcactta tgagttgctg
     3301 cctcagactc tgactttcca catgtttctt ggtcatgtaa gtttctgggg atggacccca
     3361 gatccacata cagttgctct attttaatca tgatttgcaa agtggttttt cttttttaag
     3421 ggaaaatggc aactccaaat aaaatttttt tgatgtgttt atataagagg ctacaggaag
     3481 tgctgtacat gaattccagg ccacggactg aaactgattt tcatcagcac aaactggaat
     3541 gcagggtaat gagtctgcaa ctggttaaaa tcagcctctt ctaattgatt aggcctgtct
     3601 aaaggaggct agaattacaa agtcagattc tgagtgcaag cagaccaggc ctttcctctg
     3661 ttacatttag tctgtcaatg atttgttgtc ttcggtaaat acattcaggc tgtcaatgtt
     3721 cagctctgtt catatggaac cctgttccca aattcaaacc aatttaaaat gctttgcagg
     3781 agggaggtcg gggaggtggg agcatgcaaa accgtctcac ccagatcgat gtatattagc
     3841 ccacaaaacc cctgtggcca tgcagaggta ataagaacgt gctagactct catcaagtcc
     3901 acatccctgg ataggagtag ctgttgtatc ctctttgtgc tagcacgcgt aagagctcgg
     3961 tacctatcga tagagaaatg ttctggcacc tgcacttgca ctggggacag cctattttgc
     4021 tagtttgttt tgtttcgttt tgttttgatg gagagcgtat gttagtacta tcgattcaca
     4081 caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattgcggc cgctccaagt
     4141 acctcccgta ccttaatatt acttacttat catggtagct tgggctggcg taatagcgaa
     4201 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcaaatt
     4261 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4321 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4381 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4441 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4501 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4561 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4621 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4681 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4741 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4801 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4861 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4921 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4981 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5041 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5101 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5161 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5221 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5281 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5341 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5401 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5461 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5521 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5581 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5641 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5701 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5761 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5821 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
